HPRK_BACSU
ID HPRK_BACSU Reviewed; 310 AA.
AC O34483; Q9R9E3;
DT 15-JUL-1998, integrated into UniProtKB/Swiss-Prot.
DT 23-JAN-2007, sequence version 3.
DT 03-AUG-2022, entry version 158.
DE RecName: Full=HPr kinase/phosphorylase;
DE Short=HPrK/P;
DE EC=2.7.11.-;
DE EC=2.7.4.-;
DE AltName: Full=HPr kinase/phosphatase;
DE AltName: Full=HPr(Ser) kinase/phosphorylase;
GN Name=hprK; Synonyms=ptsK, yvoB; OrderedLocusNames=BSU35000;
OS Bacillus subtilis (strain 168).
OC Bacteria; Firmicutes; Bacilli; Bacillales; Bacillaceae; Bacillus.
OX NCBI_TaxID=224308;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RA Lazarevic V., Soldo B., Rivolta C., Reynolds S., Mauel C., Karamata D.;
RT "Nucleotide sequence of the 300-304 chromosomal segment of Bacillus
RT subtilis.";
RL Submitted (NOV-1997) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=168;
RX PubMed=9384377; DOI=10.1038/36786;
RA Kunst F., Ogasawara N., Moszer I., Albertini A.M., Alloni G., Azevedo V.,
RA Bertero M.G., Bessieres P., Bolotin A., Borchert S., Borriss R.,
RA Boursier L., Brans A., Braun M., Brignell S.C., Bron S., Brouillet S.,
RA Bruschi C.V., Caldwell B., Capuano V., Carter N.M., Choi S.-K.,
RA Codani J.-J., Connerton I.F., Cummings N.J., Daniel R.A., Denizot F.,
RA Devine K.M., Duesterhoeft A., Ehrlich S.D., Emmerson P.T., Entian K.-D.,
RA Errington J., Fabret C., Ferrari E., Foulger D., Fritz C., Fujita M.,
RA Fujita Y., Fuma S., Galizzi A., Galleron N., Ghim S.-Y., Glaser P.,
RA Goffeau A., Golightly E.J., Grandi G., Guiseppi G., Guy B.J., Haga K.,
RA Haiech J., Harwood C.R., Henaut A., Hilbert H., Holsappel S., Hosono S.,
RA Hullo M.-F., Itaya M., Jones L.-M., Joris B., Karamata D., Kasahara Y.,
RA Klaerr-Blanchard M., Klein C., Kobayashi Y., Koetter P., Koningstein G.,
RA Krogh S., Kumano M., Kurita K., Lapidus A., Lardinois S., Lauber J.,
RA Lazarevic V., Lee S.-M., Levine A., Liu H., Masuda S., Mauel C.,
RA Medigue C., Medina N., Mellado R.P., Mizuno M., Moestl D., Nakai S.,
RA Noback M., Noone D., O'Reilly M., Ogawa K., Ogiwara A., Oudega B.,
RA Park S.-H., Parro V., Pohl T.M., Portetelle D., Porwollik S.,
RA Prescott A.M., Presecan E., Pujic P., Purnelle B., Rapoport G., Rey M.,
RA Reynolds S., Rieger M., Rivolta C., Rocha E., Roche B., Rose M., Sadaie Y.,
RA Sato T., Scanlan E., Schleich S., Schroeter R., Scoffone F., Sekiguchi J.,
RA Sekowska A., Seror S.J., Serror P., Shin B.-S., Soldo B., Sorokin A.,
RA Tacconi E., Takagi T., Takahashi H., Takemaru K., Takeuchi M.,
RA Tamakoshi A., Tanaka T., Terpstra P., Tognoni A., Tosato V., Uchiyama S.,
RA Vandenbol M., Vannier F., Vassarotti A., Viari A., Wambutt R., Wedler E.,
RA Wedler H., Weitzenegger T., Winters P., Wipat A., Yamamoto H., Yamane K.,
RA Yasumoto K., Yata K., Yoshida K., Yoshikawa H.-F., Zumstein E.,
RA Yoshikawa H., Danchin A.;
RT "The complete genome sequence of the Gram-positive bacterium Bacillus
RT subtilis.";
RL Nature 390:249-256(1997).
RN [3]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 242-310.
RC STRAIN=168;
RA Dartois V.A., Hoch J.A.;
RL Submitted (JUL-1996) to the EMBL/GenBank/DDBJ databases.
RN [4]
RP PHOSPHORYLATION OF CRH PROTEIN.
RC STRAIN=QB5081;
RX PubMed=9237995; DOI=10.1073/pnas.94.16.8439;
RA Galinier A., Haiech J., Kilhoffer M.-C., Jaquinod M., Stuelke J.,
RA Deutscher J., Martin-Verstraete I.;
RT "The Bacillus subtilis crh gene encodes a HPr-like protein involved in
RT carbon catabolite repression.";
RL Proc. Natl. Acad. Sci. U.S.A. 94:8439-8444(1997).
RN [5]
RP PROTEIN SEQUENCE OF 2-18, CHARACTERIZATION, AND MASS SPECTROMETRY.
RX PubMed=9570401; DOI=10.1046/j.1365-2958.1998.00747.x;
RA Reizer J., Hoischen C., Titgemeyer F., Rivolta C., Rabus R., Stuelke J.,
RA Karamata D., Saier M.H. Jr., Hillen W.;
RT "A novel protein kinase that controls carbon catabolite repression in
RT bacteria.";
RL Mol. Microbiol. 27:1157-1169(1998).
RN [6]
RP CHARACTERIZATION OF THE KINASE ACTIVITY.
RX PubMed=9465101; DOI=10.1073/pnas.95.4.1823;
RA Galinier A., Kravanja M., Engelmann R., Hengstenberg W., Kilhoffer M.-C.,
RA Deutscher J., Haiech J.;
RT "New protein kinase and protein phosphatase families mediate signal
RT transduction in bacterial catabolite repression.";
RL Proc. Natl. Acad. Sci. U.S.A. 95:1823-1828(1998).
RN [7]
RP FUNCTION.
RX PubMed=9987110; DOI=10.1046/j.1365-2958.1999.01146.x;
RA Kravanja M., Engelmann R., Dossonnet V., Bluggel M., Meyer H.E., Frank R.,
RA Galinier A., Deutscher J., Schnell N., Hengstenberg W.;
RT "The hprK gene of Enterococcus faecalis encodes a novel bifunctional
RT enzyme: the HPr kinase/phosphatase.";
RL Mol. Microbiol. 31:59-66(1999).
RN [8]
RP CHARACTERIZATION, AND COFACTOR.
RX PubMed=12009882; DOI=10.1021/bi025613y;
RA Lavergne J.-P., Jault J.-M., Galinier A.;
RT "Insights into the functioning of Bacillus subtilis HPr kinase/phosphatase:
RT affinity for its protein substrates and role of cations and phosphate.";
RL Biochemistry 41:6218-6225(2002).
RN [9]
RP INDUCTION, AND MUTAGENESIS OF GLY-148.
RX PubMed=12055300; DOI=10.1099/00221287-148-6-1805;
RA Hanson K.G., Steinhauer K., Reizer J., Hillen W., Stuelke J.;
RT "HPr kinase/phosphatase of Bacillus subtilis: expression of the gene and
RT effects of mutations on enzyme activity, growth and carbon catabolite
RT repression.";
RL Microbiology 148:1805-1811(2002).
RN [10]
RP DEPHOSPHORYLATION REACTION MECHANISM.
RX PubMed=12359880; DOI=10.1073/pnas.212410399;
RA Mijakovic I., Poncet S., Galinier A., Monedero V., Fieulaine S., Janin J.,
RA Nessler S., Marquez J.A., Scheffzek K., Hasenbein S., Hengstenberg W.,
RA Deutscher J.;
RT "Pyrophosphate-producing protein dephosphorylation by HPr
RT kinase/phosphorylase: a relic of early life?";
RL Proc. Natl. Acad. Sci. U.S.A. 99:13442-13447(2002).
RN [11]
RP CHARACTERIZATION.
RX PubMed=12411438; DOI=10.1074/jbc.m209052200;
RA Ramstroem H., Sanglier S., Leize-Wagner E., Philippe C., van Dorsselaer A.,
RA Haiech J.;
RT "Properties and regulation of the bifunctional enzyme HPr
RT kinase/phosphatase in Bacillus subtilis.";
RL J. Biol. Chem. 278:1174-1185(2003).
RN [12]
RP MUTAGENESIS OF HIS-138; GLY-153; SER-155; GLY-156; GLY-158; LYS-159;
RP SER-160 AND HIS-171.
RX PubMed=12779331; DOI=10.1021/bi034405i;
RA Pompeo F., Granet Y., Lavergne J.-P., Grangeasse C., Nessler S.,
RA Jault J.-M., Galinier A.;
RT "Regulation and mutational analysis of the HPr kinase/phosphorylase from
RT Bacillus subtilis.";
RL Biochemistry 42:6762-6771(2003).
RN [13]
RP REVIEW.
RX PubMed=15023355; DOI=10.1016/j.bbapap.2003.11.018;
RA Poncet S., Mijakovic I., Nessler S., Gueguen-Chaignon V., Chaptal V.,
RA Galinier A., Boel G., Maze A., Deutscher J.;
RT "HPr kinase/phosphorylase, a Walker motif A-containing bifunctional sensor
RT enzyme controlling catabolite repression in Gram-positive bacteria.";
RL Biochim. Biophys. Acta 1697:123-135(2004).
CC -!- FUNCTION: Catalyzes the ATP- as well as the pyrophosphate-dependent
CC phosphorylation of 'Ser-45' in HPr, a phosphocarrier protein of the
CC phosphoenolpyruvate-dependent sugar phosphotransferase system (PTS).
CC HprK/P also catalyzes the pyrophosphate-producing, inorganic phosphate-
CC dependent dephosphorylation (phosphorolysis) of seryl-phosphorylated
CC HPr (P-Ser-HPr). The two antagonistic activities of HprK/P are
CC regulated by several intracellular metabolites, which change their
CC concentration in response to the absence or presence of rapidly
CC metabolisable carbon sources (glucose, fructose, etc.) in the growth
CC medium. Also phosphorylates/dephosphorylates the HPr-like catabolite
CC repression protein crh on 'Ser-46'. Therefore, by controlling the
CC phosphorylation state of HPr and crh, HPrK/P is a sensor enzyme that
CC plays a major role in the regulation of carbon metabolism and sugar
CC transport: it mediates carbon catabolite repression (CCR), and
CC regulates PTS-catalyzed carbohydrate uptake and inducer exclusion.
CC {ECO:0000269|PubMed:9987110}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=[HPr protein]-L-serine + ATP = [HPr protein]-O-phospho-L-
CC serine + ADP + H(+); Xref=Rhea:RHEA:46600, Rhea:RHEA-COMP:11602,
CC Rhea:RHEA-COMP:11603, ChEBI:CHEBI:15378, ChEBI:CHEBI:29999,
CC ChEBI:CHEBI:30616, ChEBI:CHEBI:83421, ChEBI:CHEBI:456216;
CC -!- CATALYTIC ACTIVITY:
CC Reaction=[HPr protein]-O-phospho-L-serine + H(+) + phosphate = [HPr
CC protein]-L-serine + diphosphate; Xref=Rhea:RHEA:46604, Rhea:RHEA-
CC COMP:11602, Rhea:RHEA-COMP:11603, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:29999, ChEBI:CHEBI:33019, ChEBI:CHEBI:43474,
CC ChEBI:CHEBI:83421;
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:12009882};
CC Name=Ca(2+); Xref=ChEBI:CHEBI:29108;
CC Evidence={ECO:0000269|PubMed:12009882};
CC Note=The enzyme may harbor two different cation-binding sites, one that
CC interacts specifically with the nucleotide, and the other that is
CC involved in the binding of the protein substrate.
CC {ECO:0000269|PubMed:12009882};
CC -!- ACTIVITY REGULATION: Is active as a kinase only at high ATP
CC concentrations or at low ATP concentrations in the presence of the
CC allosteric activator fructose 1,6-bisphosphate (FBP). The
CC pyrophosphate-dependent HPr phosphorylation is not stimulated by FBP.
CC Kinase activity is inhibited by inorganic phosphate (Pi).
CC Dephosphorylation of HPr(Ser-P) by B.subtilis HPrK/P becomes prevalent
CC when the concentration of Pi increases. Thus, the kinase activity may
CC prevail under conditions of good nutrient supply, whereas the
CC phosphorylase activity is dominant if carbon and energy sources become
CC limiting.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=78 uM for HPr;
CC KM=85 uM for P-Ser-HPr;
CC KM=265 uM for phosphate;
CC KM=785 uM for pyrophosphate;
CC -!- SUBUNIT: Homohexamer.
CC -!- INTERACTION:
CC O34483; P11065: hpr; NbExp=3; IntAct=EBI-5242785, EBI-2121844;
CC O34483; O31435: ybdM; NbExp=5; IntAct=EBI-5242785, EBI-5255200;
CC O34483; P96716: ywqD; NbExp=2; IntAct=EBI-5242785, EBI-9302929;
CC -!- INDUCTION: Constitutively expressed, with or without glucose in the
CC growth medium. {ECO:0000269|PubMed:12055300}.
CC -!- DOMAIN: The Walker A ATP-binding motif also binds Pi and PPi.
CC -!- MASS SPECTROMETRY: Mass=34529; Method=MALDI;
CC Evidence={ECO:0000269|PubMed:9570401};
CC -!- MISCELLANEOUS: Both phosphorylation and phosphorolysis are carried out
CC by the same active site and suggest a common mechanism for both
CC reactions.
CC -!- MISCELLANEOUS: According to PubMed:12779331, the mutation G152A reduces
CC the kinase activity about twofold and has little effect on
CC phosphorylase activity, whereas according to PubMed:12055300, it
CC completely prevents both functions. This conflicting result may be
CC explained by limiting amounts substrates conditions used in
CC PubMed:12779331 assays.
CC -!- SIMILARITY: Belongs to the HPrK/P family. {ECO:0000305}.
CC -!- CAUTION: Was originally called HPr kinase/phosphatase, but P-Ser-HPr
CC dephosphorylation was found to follow a quite unique mechanism
CC (PubMed:12359880), in which Pi instead of H(2)O is used for the
CC nucleophilic attack on the phosphoryl group. P-Ser-HPr
CC dephosphorylation is therefore not a phosphohydrolysis but a phospho-
CC phosphorolysis reaction, and the bifunctional enzyme was dubbed HPr
CC kinase/phosphorylase. {ECO:0000305|PubMed:12359880}.
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DR EMBL; AF017113; AAC67286.1; -; Genomic_DNA.
DR EMBL; AL009126; CAB15505.1; -; Genomic_DNA.
DR EMBL; U63310; AAD09500.1; -; Genomic_DNA.
DR PIR; E70044; E70044.
DR RefSeq; NP_391380.1; NC_000964.3.
DR RefSeq; WP_003228097.1; NZ_JNCM01000033.1.
DR AlphaFoldDB; O34483; -.
DR SMR; O34483; -.
DR IntAct; O34483; 6.
DR STRING; 224308.BSU35000; -.
DR BindingDB; O34483; -.
DR ChEMBL; CHEMBL4974; -.
DR PaxDb; O34483; -.
DR PRIDE; O34483; -.
DR EnsemblBacteria; CAB15505; CAB15505; BSU_35000.
DR GeneID; 936615; -.
DR KEGG; bsu:BSU35000; -.
DR PATRIC; fig|224308.179.peg.3788; -.
DR eggNOG; COG1493; Bacteria.
DR InParanoid; O34483; -.
DR OMA; AMNNRQK; -.
DR PhylomeDB; O34483; -.
DR BioCyc; BSUB:BSU35000-MON; -.
DR SABIO-RK; O34483; -.
DR PRO; PR:O34483; -.
DR Proteomes; UP000001570; Chromosome.
DR GO; GO:0005829; C:cytosol; IBA:GO_Central.
DR GO; GO:0005524; F:ATP binding; IEA:UniProtKB-UniRule.
DR GO; GO:0000287; F:magnesium ion binding; IEA:UniProtKB-UniRule.
DR GO; GO:0000155; F:phosphorelay sensor kinase activity; IEA:InterPro.
DR GO; GO:0004674; F:protein serine/threonine kinase activity; IEA:UniProtKB-KW.
DR GO; GO:0004712; F:protein serine/threonine/tyrosine kinase activity; IEA:UniProtKB-UniRule.
DR GO; GO:0005975; P:carbohydrate metabolic process; IEA:UniProtKB-KW.
DR GO; GO:0006109; P:regulation of carbohydrate metabolic process; IEA:UniProtKB-UniRule.
DR CDD; cd01918; HprK_C; 1.
DR Gene3D; 3.40.1390.20; -; 1.
DR Gene3D; 3.40.50.300; -; 1.
DR HAMAP; MF_01249; HPr_kinase; 1.
DR InterPro; IPR003755; HPr(Ser)_kin/Pase.
DR InterPro; IPR011104; Hpr_kin/Pase_C.
DR InterPro; IPR011126; Hpr_kin/Pase_Hpr_N.
DR InterPro; IPR027417; P-loop_NTPase.
DR InterPro; IPR028979; Ser_kin/Pase_Hpr-like_N_sf.
DR PANTHER; PTHR30305:SF1; PTHR30305:SF1; 1.
DR Pfam; PF07475; Hpr_kinase_C; 1.
DR Pfam; PF02603; Hpr_kinase_N; 1.
DR SUPFAM; SSF75138; SSF75138; 1.
DR TIGRFAMs; TIGR00679; hpr-ser; 1.
PE 1: Evidence at protein level;
KW Allosteric enzyme; ATP-binding; Carbohydrate metabolism;
KW Direct protein sequencing; Kinase; Magnesium; Metal-binding;
KW Multifunctional enzyme; Nucleotide-binding; Reference proteome;
KW Serine/threonine-protein kinase; Transferase.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:9570401"
FT CHAIN 2..310
FT /note="HPr kinase/phosphorylase"
FT /id="PRO_0000058946"
FT REGION 201..210
FT /note="Important for the catalytic mechanism of both
FT phosphorylation and dephosphorylation"
FT REGION 264..269
FT /note="Important for the catalytic mechanism of
FT dephosphorylation"
FT /evidence="ECO:0000250"
FT ACT_SITE 159
FT /evidence="ECO:0000250"
FT ACT_SITE 177
FT /note="Proton acceptor; for phosphorylation activity.
FT Proton donor; for dephosphorylation activity"
FT /evidence="ECO:0000250"
FT ACT_SITE 243
FT /evidence="ECO:0000250"
FT BINDING 138
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000255"
FT BINDING 153..160
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000305"
FT BINDING 160
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000305"
FT BINDING 202
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000305"
FT MUTAGEN 138
FT /note="H->A: No effect on both kinase and phosphorylase
FT activities."
FT /evidence="ECO:0000269|PubMed:12779331"
FT MUTAGEN 148
FT /note="G->A: Loss of both kinase and phosphorylase
FT activities."
FT /evidence="ECO:0000269|PubMed:12055300"
FT MUTAGEN 153
FT /note="G->A: Loss of both kinase and phosphorylase
FT activities."
FT /evidence="ECO:0000269|PubMed:12779331"
FT MUTAGEN 155
FT /note="S->A,T: Nearly no effect on both kinase and
FT phosphorylase activities."
FT /evidence="ECO:0000269|PubMed:12779331"
FT MUTAGEN 155
FT /note="S->C: Loss of both kinase and phosphorylase
FT activities."
FT /evidence="ECO:0000269|PubMed:12779331"
FT MUTAGEN 156
FT /note="G->A: Loss of both kinase and phosphorylase
FT activities."
FT /evidence="ECO:0000269|PubMed:12779331"
FT MUTAGEN 158
FT /note="G->A: Loss of both kinase and phosphorylase
FT activities."
FT /evidence="ECO:0000269|PubMed:12779331"
FT MUTAGEN 159
FT /note="K->R: Slight reduction in both kinase and
FT phosphorylase activities."
FT /evidence="ECO:0000269|PubMed:12779331"
FT MUTAGEN 160
FT /note="S->A: Loss of both kinase and phosphorylase
FT activities."
FT /evidence="ECO:0000269|PubMed:12779331"
FT MUTAGEN 160
FT /note="S->T: Nearly no effect on both kinase and
FT phosphorylase activities."
FT /evidence="ECO:0000269|PubMed:12779331"
FT MUTAGEN 171
FT /note="H->R: Loss of both kinase and phosphorylase
FT activities."
FT /evidence="ECO:0000269|PubMed:12779331"
FT MUTAGEN 202
FT /note="E->A: No effect on ATP-dependent kinase activity,
FT but loss of both PPi-kinase and phosphorylase activities."
FT MUTAGEN 204
FT /note="R->A: Reduced kinase activity and loss of
FT phosphorylase activity."
FT MUTAGEN 205
FT /note="G->A: No effect on ATP-dependent kinase activity,
FT but loss of both PPi-kinase and phosphorylase activities."
FT MUTAGEN 207
FT /note="G->A: Reduced kinase activity and loss of
FT phosphorylase activity. No effect on FBP binding."
SQ SEQUENCE 310 AA; 34702 MW; C44ABD220881EC31 CRC64;
MAKVRTKDVM EQFNLELISG EEGINRPITM SDLSRPGIEI AGYFTYYPRE RVQLLGKTEL
SFFEQLPEEE KKQRMDSLCT DVTPAIILSR DMPIPQELID ASEKNGVPVL RSPLKTTRLS
SRLTNFLESR LAPTTAIHGV LVDIYGVGVL ITGKSGVGKS ETALELVKRG HRLVADDCVE
IRQEDQDTLV GNAPELIEHL LEIRGLGIIN VMTLFGAGAV RSNKRITIVM NLELWEQGKQ
YDRLGLEEET MKIIDTEITK LTIPVRPGRN LAVIIEVAAM NFRLKRMGLN AAEQFTNKLA
DVIEDGEQEE
